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hIL-1b QKit (1 Kit)
品牌： R&D Systems
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Sample Values
Serum/Plasma - Forty serum and plasma samples from apparently healthy volunteers were evaluated for the presence of human IL-1 beta in this assay. No medical histories were available for the donors used in this study. All samples measured less than the lowest IL-1 beta standard, 3.9 pg/mL.Cell Culture Supernates - Human peripheral blood mononuclear cells (1 x 106 cells/mL) were cultured in RPMI supplemented with 10% fetal bovine serum, 50 μM beta -mercaptoethanol, 2 mM L-glutamine, 100 U/mL penicillin, and 100 μg/mL streptomycin sulfate. Cells were stimulated with the agents listed in the table below. Aliquots of the cell culture supernate were removed on days 1, 3, and 5 and assayed for levels of human IL-1 beta.
| Stimulant | Day 1 (pg/mL) | Day 3 (pg/mL) | Day 5 (pg/mL) |
| 10 μg/mL PHA | 2185 | 2004 | 2383 |
| 10 μg/mL PHA+10 ng/mL rhIL-2 | 1938 | 1973 | 2839 |
| 50 ng/mL PMA | 1767 | 1027 | 1159 |
| 50 ng/mL LPS | 4158 | 2145 | 1308 |
Recovery
The recovery of IL-1 beta spiked to different levels throughout the range of the assay in various matrices was evaluated.
| Sample Type | Average % Recovery | Range % |
|---|---|---|
| Cell Culture Media (n=4) | 97 | 80-111 |
| Citrate Plasma (n=10) | 93 | 83-110 |
| EDTA Plasma (n=10) | 86 | 81-100 |
| Heparin Plasma (n=10) | 82 | 76-100 |
| Serum (n=10) | 95 | 87-110 |
Linearity
To assess linearity of the assay, the following biological samples containing, or spiked with, high concentrations of IL-1 beta were diluted with the appropriate Calibrator Diluent and then assayed.
Scientific Data
Assay Procedure
Refer to the product Bring all reagents and samples to room temperature before use. It is recommended that all samples, standards, and controls be assayed in duplicate.
- Prepare all reagents, standard dilutions, and samples as directed in the product insert.
- Remove excess microplate strips from the plate frame, return them to the foil pouch containing the desiccant pack, and reseal.
- For Serum & Plasma Samples: Add 50 µL of Assay Diluent to each well.
- Add 200 µL of Standard, control, or sample to each well. Cover with a plate sealer, and incubate at room temperature for 2 hours.
- Aspirate each well and wash, repeating the process twice for a total of 3 washes.
- Add 200 µL of Conjugate to each well.
- For Cell Culture Supernate Samples: Cover with a new plate sealer, and incubate at room temperature for 1 hour.
For Serum & Plasma Samples: Cover with a new plate sealer, and incubate at room temperature for 2 hours. - Aspirate and wash 3 times.
- Add 200 µL Substrate Solution to each well. Incubate at room temperature for 20 minutes. PROTECT FROM LIGHT.
- Add 50 µL of Stop Solution to each well. Read at 450 nm within 30 minutes. Set wavelength correction to 540 nm or 570 nm.
50 µL Assay Diluent for Serum & Plasma Samples Only
200 µL Standard, Control, or Sample
200 µL Conjugate
200 µL Substrate Solution
50 µL Stop Solution
Human IL-1 beta/IL-1F2 Quantikine ELISA Kit Summary
Assay Type
Solid Phase Sandwich ELISA
Format
96-well strip plate
Assay Length
3.5 hours or 4.5 hours
Sample Type & Volume Required Per Well
Cell Culture Supernates (200 uL), Serum (200 uL), EDTA Plasma (200 uL), Heparin Plasma (200 uL), Citrate Plasma (200 uL)
Sensitivity
1 pg/mL
Assay Range
3.9 - 250 pg/mL (Cell Culture Supernates, Serum, EDTA Plasma, Heparin Plasma, Citrate Plasma)
Specificity
Natural and recombinant human IL-1 beta
Cross-reactivity
< 0.5% cross-reactivity observed with available related molecules.< 50% cross-species reactivity observed with species tested.
Interference
Interference observed with 1 or more available related molecules.
背景
别名
catabolin,IL1 beta,IL-1 beta,IL-1,IL1B,IL-1b,IL1-BETA,IL-1F2,IL1F2IL-1 beta,interleukin 1, beta,interleukin-1 beta,preinterleukin 1 beta,pro-interleukin-1-beta,Interleukin 1 beta
背景
Background: IL-1 beta/IL-1F2
The Interleukin 1 (IL-1) family of proteins consists of the classic members IL-1 alpha, IL-1 beta, and IL-1ra, plus IL-18, IL-33 and IL-1F5-F10. IL-1 alpha and IL-1 beta bind to the same cell surface receptors and share biological functions. IL-1 is not produced by unstimulated cells of healthy individuals with the exception of skin keratinocytes, some epithelial cells, and certain cells of the central nervous system. However, in response to inflammatory agents, infections, or microbial endotoxins, a dramatic increase in the production of IL-1 by macrophages and various other cell types is observed. IL-1 beta plays a central role in immune and inflammatory responses, bone remodeling, fever, carbohydrate metabolism, and GH/IGF-I physiology. Inappropriate or prolonged production of IL-1 has been implicated in a variety of pathological conditions including sepsis, rheumatoid arthritis, inflammatory bowel disease, acute and chronic myelogenous leukemia, insulin dependent diabetes mellitus, atherosclerosis, neuronal injury, and aging-related diseases.
IL-1 alpha and IL-1 beta are structurally related polypeptides that show approximately 25% homology at the amino acid level. Both are synthesized as 31 kDa precursors that are subsequently cleaved into mature proteins of approximately 17.5 kDa. Cleavage of the IL-1 beta precursor by Caspase-1/ICE is a key step in the inflammatory response. Neither IL-1 alpha nor IL-1 beta contains a typical hydrophobic signal peptide, but evidence suggests that these factors can be secreted by non-classical pathways. A portion of unprocessed IL-1 alpha can be presented on the cell membrane and may retain biological activity. The precursor form of IL-1 beta, unlike the IL-1 alpha precursor, shows little or no biological activity in comparison to the processed form. Both unprocessed and mature forms of IL-1 beta are exported from the cell.
IL-1 alpha and IL-1 beta exert their effects through immunoglobulin superfamily receptors that additionally bind IL-1ra. The 80 kDa transmembrane type I receptor (IL-1 RI) is expressed on T cells, fibroblasts, keratinocytes, endothelial cells, synovial lining cells, chondrocytes, and hepatocytes. The 68 kDa transmembrane type II receptor (IL-1 RII) is expressed on B cells, neutrophils, and bone marrow cells. The two IL-1 receptor types show approximately 28% homology in their extracellular domains but differ significantly in that the type II receptor has a cytoplasmic domain of only 29 amino acids (aa), whereas the type I receptor has a 213 aa cytoplasmic domain. IL-1 RII does not appear to signal in response to IL-1 and may function as a decoy receptor that attenuates IL-1 function. The IL-1 receptor accessory protein (IL-1 RAcP) associates with IL-1 RI and is required for IL-1 RI signal transduction. IL-1ra is a secreted molecule that functions as a competitive inhibitor of IL-1. Soluble forms of both IL-1 RI and IL-1 RII have been detected in human plasma, synovial fluids, and the conditioned media of several human cell lines. In addition, IL-1 binding proteins that resemble soluble IL-1 RII are encoded by vaccinia and cowpox viruses.
Long Name:
Interleukin 1 beta
Entrez Gene IDs:
3553 (Human); 16176 (Mouse); 24494 (Rat); 397122 (Porcine); 403974 (Canine); 100034237 (Equine); 100135556 (Guinea Pig)
Alternate Names:
catabolin; IL1 beta; IL-1 beta; IL-1; IL1B; IL-1b; IL1-BETA; IL-1F2; IL1F2IL-1 beta; interleukin 1, beta; interleukin-1 beta; preinterleukin 1 beta; pro-interleukin-1-beta
研究领域
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